The Impact of Mulberry Fruit Extracts on Skin Aging: Comprehensive Assessments of Their Role in Hindering Glycation and Enhancing Antioxidant Functions, Both in Lab Tests and Real-World Applications.

Posted on January 3, 2024  •  8 minutes  • 1588 words  • Other languages:  Русский

Introduction

Mulberry, a versatile agricultural product, is recognized for its health-promoting qualities and is a subject of extensive research as a beneficial health food. This fruit, along with its leaves, twigs, and roots, is packed with active compounds. Analysis has revealed that mulberry fruit primarily comprises two key functional components: polyphenols and polysaccharides, known for their antioxidant, anti-inflammatory, anti-cancer, memory-enhancing, and neuroprotective effects. Mulberry fruit extracts are particularly noted for their phenolic substances, which boost antioxidant enzyme activity and help slow down memory loss in aging by aiding the removal of amyloid-β. Moreover, mulberries are abundant in water-soluble bioactive components known as anthocyanins, which have been shown to decelerate aging and mitigate age-related conditions.

The aging process brings various degenerative diseases, and the skin, being the body’s largest organ, is vulnerable to internal oxidative stress from numerous biochemical processes, including oxidation and glycosylation. Advanced Glycation End-products (AGEs) are complex compounds formed through the reaction of reducing sugars with protein amino groups, resulting in Schiff bases and rearrangements. The accumulation of AGEs leads to protein denaturation, oxidative stress, and inflammation, playing a significant role in the development of several skin diseases, including aging-related skin issues. Skin glycosylation affects the dermal matrix and long-lived proteins, reducing tissue elasticity. With aging, the buildup of AGEs in the skin leads to issues like dullness and wrinkles. Skin aging is primarily driven by a shift in oxidative homeostasis and the diminished activity of keratin-producing cells in aged skin. Studies also highlight that natural polyphenols can effectively slow down and prevent skin aging.

The anti-glycation effects of Mulberry Fruit Extracts (MFEs) are yet to be fully understood. The current study involved preparing extracts from various mulberry sources to explore the potential of phenolic substances in MFEs. The study examined the anti-aging, anti-glycation, and antioxidant properties of MFEs using the HaCaT cell line and an aging mouse model induced by D-galactose. This research provides a basis for the future application of mulberry fruit extract in the food and cosmetic industries.

Experiments

1. Materials and Reagents:

   • Mulberry (Morus alba L.) was sourced from a specific company.
   • Various chemicals and reagents were purchased, including BSA, MGO, and aminoguanidine.
   • Various cell culture materials and equipment were obtained from different suppliers.

2. Preparation of Mulberry Extract:

   • Three types of raw materials were used: air-dried mulberry fruits (DMF), mulberry fruit juice (MJ), and mulberry fruit pomace (MFP).
   • Three extraction methods were applied: water extraction, Viscozyme® L -assisted extraction, and Pectinex® XXL -assisted extraction.
   • The yield rate of each extraction was calculated.

3. Phenolic Compounds Analysis:

   • Utilized UPLC-MS/MS for analysis.
   • Found differences in active substances in the extracts based on the extraction method and materials.

4. Determination of Antioxidant Activity:

   • The ABTS scavenging capacity of the extracts was determined.
   • This involved measuring the Trolox equivalent antioxidant capacity.

5. In vitro Anti-glycation Assay:

   • Two models were used: MGO-BSA and Fructose-BSA.
   • Evaluated the anti-glycation ability of the mulberry extracts using these models.

6. Cell Viability and Oxidative Stress Determination:

   • Used HaCaT cells to assess cell viability and oxidative stress.
   • Investigated the protective effects of the extracts against MGO-induced cell injury.

7. Cell Apoptosis Analysis:

   • Analyzed the effect of the extracts on apoptosis in HaCaT cells.

8. Animal Treatments and Skin Index Measurement:

   • Conducted on D-galactose-induced aging mice.
   • Measured various skin parameters, including moisture, HA, HYP, and AGEs.

9. Serum Oxidative Stress Determination:

   • Analyzed the effect of the extracts on oxidative stress parameters in the serum of aging mice.

10. Statistical Analysis:

    • Data were expressed as mean ± SD and analyzed using one-way ANOVA and other statistical tests.

The experiments were comprehensive, exploring the effects of mulberry fruit extracts on anti-glycation, antioxidant activities, and their potential protective effects against skin aging both in vitro (using cell lines) and in vivo (using a mouse model). The study meticulously evaluated various extraction methods, their yield, and the bioactivity of the resulting extracts, highlighting the potential of mulberry fruit extracts in skincare and anti-aging applications.

Results

Yield Rate of Different Mulberry Fruit Extracts (MFEs)

Different raw materials and extraction methods showed significant differences in yield rates. The highest yield was obtained from direct hot water extraction.

Total Phenolic Content in Extracts

The total phenolic content varied among the different extracts, with air-dried mulberry fruits (DMF) showing the highest content.

Analysis of Phenolic Components in Extracts

UPLC-MS/MS analysis showed variations in phenolic components among different extracts. The analysis revealed higher numbers of unique compounds in mulberry juice (MJ) samples compared to DMF and mulberry fruit pomace (MFP).

ABTS Free Radical Scavenging Ability of Extracts

The extracts showed significant variations in ABTS free radical scavenging ability, with MFP extracts demonstrating the highest scavenging ability.

In Vitro Anti-glycation Effect of Extracts

The extracts showed significant anti-glycation effects, with Pectinex® XXL-assisted extracts showing stronger effects in the BSA-fructose system. In the MGO-BSA system, DMF-W, DMF-P, and MFP-V exhibited superior anti-glycation ability.

Protection Effects of MFEs on MGO-induced HaCaT Injury

Some extracts, especially DMF-P, showed significant protective effects against MGO-induced injury in HaCaT cells.

Effect of MFEs on the Level of Oxidative Stress in HaCaT Cells

Treatment with MFEs significantly reduced Malondialdehyde (MDA) levels and increased Superoxide Dismutase (SOD) activity in HaCaT cells, indicating a reduction in oxidative stress.

Effect of MFEs on MGO-induced Apoptosis of HaCaT Cells

MFEs, particularly DMF-P, effectively reduced apoptosis in HaCaT cells induced by MGO.

Effects of MFEs on Relevant Indicators in the Skin of Aging Mice

MFEs improved skin moisture, increased hyaluronic acid and hydroxyproline content, and reduced AGEs in D-galactose-induced aging mice, indicating anti-aging effects.

Effects of MFEs on Oxidative Stress in Aging Model Mice

MFEs enhanced serum GSH-Px content and SOD activity while reducing serum MDA levels in D-galactose-induced aging mice, suggesting a reduction in oxidative stress.

Overall, the study demonstrates that mulberry fruit extracts, particularly those extracted using Pectinex® XXL, have significant anti-glycation and antioxidant effects. They show potential in protecting against skin aging, reducing oxidative stress and apoptosis in vitro, and improving skin condition in an aging mice model.

Conclusion

In this research, various extracts of mulberry fruit (MFEs) were evaluated for their capacity to counteract glycation both in laboratory settings and through animal studies. The outcomes showed a substantial reduction in Advanced Glycation End products (AGEs), with the fructose-BSA model showing over 40% decrease and the MGO-BSA model exhibiting an impressive 73% reduction. The cellular experiments indicated that MFEs mitigated the oxidative stress and cell death caused by MGO in HaCaT cells. Analysis of the extracts' composition pointed towards phenolic elements as key contributors to their beneficial effects. Furthermore, experiments on mice artificially aged with D-galactose revealed that DMF-W and DMF-P significantly improved skin hydration, hyaluronic acid, and hydroxyproline levels, enhancing skin quality and lowering oxidative stress. These extracts also reduced AGEs in the skin of these aged mice. These findings suggest that DMF-W and DMF-P, in particular, could be effective in slowing down skin aging processes and associated oxidative harm in mice, pointing to the potential of MFEs, especially DMF variants, in combating skin aging through the inhibition of glycation and oxidative stress, thereby exerting anti-aging effects on the skin.

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